Modulation of ethylene and ROS-scavenging enzymes by multifarious plant growth-promoting endophytes in tomato (Solanum lycopersicum) plants to combat Xanthomonas -induced stress.

The purpose of the current study was to explore root endophytes- Priestia megaterium T3 and Bacillus cereus T4 from Moringa olefiera for the suppression of leaf spot disease in tomato plants challenged with Xanthomonas vesicatoria. Both strains had plant growth-stimulating characteristics including auxin production, solubilization of inorganic phosphate and zinc complexes, and production of ammonia, siderophore, as well as hydrolytic enzymes. An agar well diffusion and fluorescence viability assay have validated the antibacterial effect of the cell-free culture supernatant of strains T3 and T4. Liquid chromatography-mass spectrometry (LC-MS) profiling has identified the secondary metabolites in the cell-free supernatant of strains T3 and T4. The bio-priming of tomato seeds with a consortium of T3 and T4 strains has significantly declined ethylene (by 0.61-fold) and hydrogen peroxide (H2O2, 0.64-fold) concentration thus, maintaining a lower content of ROS-induced malondialdehyde (MDA, 0.91-fold) as compared to control counterparts. Consequently, the leaf spot disease severity was reduced by ∼70% in consortium-treated tomato plants in contrast to their pathogen-challenged control. The consortia (T3+T4) treatment has facilitated induced systemic resistance by enhancing enzymatic activities of phenylalanine ammonia-lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO), catalase (CAT), and ascorbate oxidase (AO) to detoxify the excessive Xanthomonas-induced ROS accumulation in tomato plants. Conclusively, bacterial endophytes modulate X. vesicatoria-induced ROS response and ethylene levels in tomato plants. The current findings indicate that plant growth-promoting endophytic bacterial strains hold the potential to sustainably enhance plant growth and suppress bacterial leaf spot disease in tomato plants.

Uncovering the effect of Moringa oleifera Lam. leaf addition to Fuzhuan Brick Tea on sensory properties, volatile profiles and anti-obesity activity.

As a nutritious plant with valuable potential, the Moringa oleifera Lam. leaf addition to Fuzhuan Brick Tea (FBT) for co-fermentation is an industrial innovation and a new route to make full use of Moringa oleifera Lam. leaves. However, the sensory properties, volatile profiles and anti-obesity activity of Fuzhuan Brick (Moringa oleifera Lam.) tea (MFBT) are still unknown. The results demonstrated that MFBT has richer and more complex smell and taste, better color and higher overall acceptance scores. In total, 57 volatile flavor compounds, consisting of 3 acids, 16 hydrocarbons, 5 esters, 8 ketones, 13 aldehydes, 6 alcohols and others, were identified using HS-SPME-GC-MS. The characteristic odor components in MFBT were 3-buten-2-one, 4-(2,6,6-trimethyl-1-cyclohexen-1-yl)- and 1-cyclohexene-1-carboxaldehyde, 2,6,6-trimethyl-, which gave it a floral, woody, sweet, herbal and fruity aroma. 2-Octenal, (E) contributed significantly to the aroma of FBT, which could impart fresh, fatty and green aromas. In addition, MFBT could better regulate lipid accumulation, glucose tolerance, insulin tolerance and inflammation response more effectively than FBT. The mechanism is that MFBT could better regulate the dysbiosis of gut microbiota induced by HFFD, mainly increasing the abundance of beneficial bacteria such as SCFA-producing bacteria (Bacteroidetes, Lactobacillaceae, Bacteroidales_S24-7_group and Clostridiaceae_1) and decreasing the abundance of harmful bacteria such as pro-inflammatory/obesity and metabolic syndrome-related bacteria (Proteobacteria, Deferribacteres, Desulfovibrio, Catenibacterium and Helicobacter), which in turn increased feces short-chain fatty acids and lowered circulating lipopolysaccharides. These results suggested that co-fermentation with Moringa oleifera Lam. leaf could significantly improve the quality and enhance the anti-obesity effect of FBT.

Antiproliferative and Oxidative Damage Protection Activities of Endophytic Fungi Aspergillus fumigatus and Chaetomium globosum from Moringa oleifera Lam.

The current study was conducted to evaluate the antiproliferative and oxidative damage protection potential of endophytic fungi Aspergillus fumigatus and Chaetomium globosum isolated from Moringa oleifera. The chloroformic extract (CE) of both the fungi showed dose dependent antiproliferative activity against human prostate adenocarcinoma (PC-3) cell line with (IC50) value of 0.055 mg/ml and 0.008 mg/ml, respectively. Further, CE of both the fungi was studied for their ability to induce apoptosis in PC-3 cell line. Various deformities in the cancerous cells treated with CE of both the fungi have been observed by confocal microscopy which indicates the cell death by apoptosis. Further apoptosis inducing ability of CE of both the fungi was observed using various flow cytometric studies. The chloroformic extract of both the fungi showed slight increase in the level of reactive oxygen species to induce apoptosis. It also showed arrest of cancerous cells at G0/G1 phase of cell cycle to induce apoptosis. The externalization of phosphatidylserine (PS) to induce apoptosis was also observed when analysed using Annexin V-FITC/PI double staining assay where the CE of A. fumigatus and C. globosum showed the total apoptosis of 94.2% and 90.3%, respectively, at the highest tested concentration of GI70. The CE of both the fungi further showed the protective behaviour for plasmid DNA pBR322, when tested for their effect against the oxidative stress caused by the Fenton's reagent. Thus, the studies demonstrated a good antiproliferative and oxidative damage protection potential of the endophytic fungi.

Bioactive potential of endophytic fungus Chaetomium globosum and GC-MS analysis of its responsible components.

The recent exploration of various medicinal plants for bioactive potential has led to the growing interest to explore their endophytes for such bioactive potential which may turn out to be better option than the plants. In the present study, Chaetomium globosum, an endophytic fungus isolated from Moringa oleifera Lam has been explored for its various biological activities. The chloroformic extract of C. globosum showed good antimutagenicity against the reactive carcinogenic mutagen, 2-aminofluorene (2-AF) in Ames test. The antiproliferative activity against various cell lines such as HCT-15, HeLa and U87-MG was found to be dose dependent and the viability reduced to 9.26%, 15.7% and 16.3%, respectively. Further, the chloroformic fungal extract was investigated for free radical scavenging activity using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethyl-benzthiazolin-6-sulfonic acid) assay which showed the IC50 value of 45.16 µg/ml and 50.55 µg/ml, respectively. The fungal extract also showed good ferric reducing power. Total phenolic and flavonoid content was found to be in linear relationship with the antioxidant potential of the fungal extract. High performance liquid chromatography showed the presence of phenolics which may help to combat the free radicals. The presence of various bioactive compounds was analysed by GC-MS which endorsed Chaetomium globosum to be a promising candidate for drug development.

Antibiofilm, antiproliferative, antioxidant and antimutagenic activities of an endophytic fungus Aspergillus fumigatus from Moringa oleifera.

An endophytic fungus Aspergillus fumigatus isolated from Moringa oleifera has been evaluated for its various bioactivities. The chloroformic fungal extract exhibited a good antimicrobial as well as antibiofilm activity against various pathogenic microorganisms. It also demonstrated a good antimutagenicity against the reactive carcinogenic ester generating mutagen, 2-aminofluorene (2-AF) with IC50 values of 0.52 mg ml-1 and 0.36 mg ml-1 in case of co-incubation and pre-incubation, respectively. The antiprolifertive activity against different cancer cell lines; such as HCT-15, HeLa A549 and U87-MG showed the IC50 values of 0.061, 0.065 and 0.072 mg ml-1, respectively. The antioxidant activity of fungal extract has been assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethyl-benzthiazolin-6-sulfonicacid) (ABTS) methods with IC50 values of 40.07 µg and 54.28 µg, respectively. Total phenolics and flavonoid contents have been also determined. Ultra-high performance liquid chromatography (UPLC) of fungal extract revealed the presence of various phenolic compounds (caffeic acid, rutin, ellagic acid, quercetin and kaempferol). Further an attempt has been made to purify the bioactive compounds by column chromatography and GC-MS analysis. The above studies demonstrated a good bioactive potential of endophytic fungus Aspergillus fumigatus and shows the pharmacological importance of an endophytic fungus and justify the need to carry out further studies.

Antimicrobial activity of endophytic fungi from the medicinal plants Mammea americana (Calophyllaceae) and Moringa oleifera (Moringaceae) / [Actividad antimicrobiana de hongos endófitos de las plantas medicinales Mammea americana (Calophyllaceae) y Moringa oleifera (Moringaceae)].

Introduction: Infectious diseases represent one of the leading causes of death worldwide. Considering the growing global challenge of antimicrobial resistance, research into new sources of potentially effective antimicrobial agents from natural origins is of great importance for world health. Objective: To evaluate the antimicrobial activity of endophytic fungi from Mammea americana and Moringa oleifera upon Staphylococcus aureus (ATCC 29213), S. aureus (resistant strain USb003), Escherichia coli (ATCC 25922), and E. coli (resistant strain USb007). Materials and methods: We isolated endophytic fungi from the leaves, seeds, and stems of the two plants under study. We evaluated their antimicrobial activity through the formation of sensitivity haloes in dual tests in vitro, as well as in trials using crude ethanolic extracts from the endophytes. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and cytotoxicity o the substances were analyzed. Results: Three ethanolic extracts of Penicillium sp., Cladosporium (001), and Cladosporium (002) exhibited the greatest inhibition halos in sensitive and resistant strains of E. coli and S. aureus. The MIC and CBM found were statistically significant (p≤0.05) compared with the gentamicin control. Furthermore, the cytotoxicity test results of CC50>1,000 demonstrated that the endophytic fungi studied exhibit bactericidal characteristics without causing unintended damage. Conclusion: The endophytic fungi M. oleifera and M. americana represent a source of active secondary metabolites with antimicrobial and non-toxic properties. In light of these findings, further research should proceed with chemical identification of the compounds and the study of their mechanisms of action, especially given the paucity of current scientific knowledge concerning the isolation of endophytes in these plants.

Introducción. Las enfermedades infecciosas son una causa importante de muertes en el mundo. La resistencia antimicrobiana es un problema global, por lo que es conveniente la investigación de nuevas fuentes de agentes antimicrobianos de origen natural potencialmente efectivos. Objetivo. Evaluar la actividad antimicrobiana de hongos endófitos de Mammea americana y Moringa oleifera en la cepa sensible (ATCC 29213) y en la cepa resistente (USb003) de Staphylococcus aureus, así como en la cepa sensible (ATCC 25922) y la cepa resistente (USb007) de Escherichia coli. Materiales y métodos. Se aislaron 14 hongos endófitos de las hojas, semillas y tallos de las dos plantas en estudio. Se evaluó su actividad antimicrobiana mediante la formación de halos de sensibilidad por ensayo dual in vitro y pruebas con extractos etanólicos crudos provenientes de los endófitos a los que se les evaluó la concentración mínima inhibitoria (CMI), la concentración bactericida mínima (CBM) y la citotoxicidad. Resultados. Tres extractos etanólicos de Penicillium sp., Cladosporium sp. (001) y Cladosporium sp. (002) presentaron mayores halos de inhibición en cepas sensibles y resistentes de E. coli y S. aureus. La CMI y la CBM halladas fueron estadísticamente significativas (p≤0,05), comparadas con el control de gentamicina. Las pruebas de citotoxicidad (concentración citotóxica, CC50>1.000) demostraron que los hongos endófitos poseen características bactericidas y no ocasionan daño alguno. Conclusión. Se halló una fuente de metabolitos secundarios activos con propiedades antimicrobianas y no tóxicas en los hongos endófitos de M. oleifera y M. americana; estos hallazgos son importantes para continuar con la identificación química de los compuestos y el estudio de sus mecanismos de acción en estas plantas en las que el aislamiento de endófitos ha sido escaso.

Dynamics of Bacterial Community and Fermentation Quality during Ensiling of Wilted and Unwilted Moringa oleifera Leaf Silage with or without Lactic Acid Bacterial Inoculants.

To investigate the effects of wilting and lactic acid bacterial inoculants on the fermentation quality and bacterial community of Moringa oleifera leaf silage, fresh and wilted M. oleifera leaves were ensiled with or without Lactobacillus farciminis LF or Lactococcus lactis LL for 1, 7, 14, 30, and 60 days. The results showed that wilting, inoculants, and their interaction exerted significant (P < 0.05) effects on the fermentation characteristics covering dry matter loss, pH value, lactic acid bacterial number, the ratio of lactic acid to acetic acid, and the relative abundances of bacteria, like for species of Lactobacillus, Lactococcus, Pediococcus, Enterococcus, Leuconostoc, and Enterobacter Both LF and LL improved the fermentation quality of wilted and unwilted M. oleifera leaf silage by accelerating lactic acid production and pH decline, decreasing dry matter loss, and inhibiting yeast and coliform bacterial growth through the whole fermentation process. During ensiling, the abundances of Lactococcus, Enterococcus, and Leuconostoc spp. increased from day 1 to day 7 and then declined sharply from day 7 to day 14. Members of these genera and Enterobacter were inhibited, whereas Lactobacillus spp. were enhanced by these two lactic acid bacterial inoculants. The relative abundances of Enterobacter, Enterococcus, and Pediococcus spp. in inoculated silages were relatively low during the whole ensiling process. A lower abundance of Enterobacter spp. was observed in wilted silages than in unwilted silages. In summary, wilting and lactic acid bacterial inoculants had an influence on bacterial community and the fermentation process; LF and LL improved the fermentation quality of wilted and unwilted M. oleifera leaf silage.IMPORTANCEMoringa oleifera leaf is a high-quality feed source for livestock and is increasingly used all over the world. Ensiling might be an effective method for preservation of the leaves. In the practice of silage making, lactic acid bacterial inoculants and wilting are commonly used to improve nutrition preservation. Monitoring the changes in a bacterial community during fermentation gives an insight into understanding and improving the ensiling process. Our results suggest that wilting and lactic acid bacterial inoculants had an influence on the bacterial community and fermentation process of M. oleifera leaf silage. Wilting showed positive effects on silage fermentation by decreasing the abundance of Enterobacter spp., while LF and LL improved the fermentation quality by inhibiting Enterobacter spp. and enhancing Lactobacillus spp. Both LF and LL accelerated the ensiling process from cocci (like Lactococcus, Enterococcus, and Leuconostoc spp.) to lactobacilli.

Bioaccessibility, antioxidant activity and modulation effect on gut microbiota of bioactive compounds from Moringa oleifera Lam. leaves during digestion and fermentation in vitro.

This study aims to investigate the bioaccessibility, bioactivity and gut microbiota modulation effect of Moringa oleifera Lam. leaves after in vitro gastrointestinal digestion and colonic fermentation. A total of 24.95 ± 0.91 mg GAE per g DM and 21.66 ± 1.41 mg RE per g DM of phenolics and flavonoids were released during digestion. The HPLC-ESI-TOF/MS analysis showed that the major phenolic compound 6,8-di-C-glucosylapigenin was released during oral digestion, catechin during gastric digestion, and quercetin-3-O-ß-d-glucoside and ferulic acid during small intestine digestion. The antioxidant activity of Moringa oleifera Lam. leaves during the whole digestion process was in line with the release of phenolics and flavonoids. In addition, the fermentation of Moringa oleifera Lam. leaves induced the production of acetic, propionic, and n-butyric acids leading to a decrease in pH, as well as the growth of beneficial colonic bacteria. The results suggested that Moringa oleifera Lam. leaves are a promising candidate for healthy food.

Antimicrobial Potential of Fungal Endophytes from Moringa oleifera.

The present study was aimed to isolate the endophytic fungi having antimicrobial potential from Moringa oleifera. Out of the active isolates, the endophytic fungal isolate DSE 17 obtained from the bark of the plant was selected for further studies and identified as Aspergillus fumigatus. The classical method for optimization strategy revealed inoculum size of four discs in Czapek dox's medium at a temperature of 25 °C and pH 7 with the incubation period of 6 days to be the best. Sucrose as carbon source (1%) and sodium nitrate as nitrogen source (0.2%) were found to be the best for antimicrobial activity. Response surface methodology was effective in optimizing the selected medium components in Plackett-Burman design, i.e. magnesium sulphate, dipotassium phosphate and sodium nitrate, which resulted in increase in antimicrobial activity by 1.7-fold. Chloroform was found to be the best extractant amongst different solvents. The minimum inhibitory concentration (MIC) values of the chloroformic extract ranged from 0.05 to 0.5 mg/ml, and the viable cell count studies revealed it to be bactericidal in its nature. The post-antibiotic effect (PAE) of the chloroformic extracts ranged from 2 to 20 h. Ames mutagenicity testing and MTT assay revealed the crude extract neither cytotoxic nor mutagenic, thus showing it to be biosafe. Thus, the study suggests that endophytes from this miracle plant could be potential source for the production of broad-spectrum antimicrobial compound/s.

Fermented Moringa oleifera Decreases Hepatic Adiposity and Ameliorates Glucose Intolerance in High-Fat Diet-Induced Obese Mice.

Metabolic diseases, such as glucose intolerance and nonalcoholic fatty-liver disease (NAFLD), are primary risk factors for life-threatening conditions such as diabetes, heart attack, stroke, and hepatic cancer. Extracts from the tropical tree Moringa oleifera show antidiabetic, antioxidant, anti-inflammatory, and anticancer effects. Fermentation can further improve the safety and nutritional value of certain foods. We investigated the efficacy of fermented M. oleifera extract (FM) against high-fat diet (HFD)-induced glucose intolerance and hepatic lipid accumulation and investigated the underlying mechanisms by analyzing expression of proteins and genes involved in glucose and lipid regulation. C57BL/6 mice were fed with normal chow diet (ND) or HFD supplemented with distilled water (DW, control), nonfermented M. oleifera extract (NFM), or FM for 10 weeks. Although body weights were similar among HFD-fed treatment groups, liver weight was decreased, and glucose tolerance test (GTT) results improved in the FM group compared with DW and NFM groups. Hepatic lipid accumulation was also lower in the FM group, and expressions of genes involved in liver lipid metabolism were upregulated. In addition, HFD-induced endoplasmic reticulum (ER) stress, oxidative stress, and lipotoxicity in quadriceps muscles were decreased by FM. Finally, proinflammatory cytokine mRNA expression was decreased by FM in the liver, epididymal adipose tissue, and quadriceps of HFD-fed mice. FMs may decrease glucose intolerance and NAFLD under HFD-induced obesity by decreasing ER stress, oxidative stress, and inflammation.

Detección de fusarium semitectum en vainas de plantas de moringa (moringa oleifera lam) / Detection of fusarium semitectum from pods of moringa plants (moringa oleífera lam)

Se analizaron muestras de Moringa oleifera procedentes de la provincia Habana, Cuba con el objetivo de determinar el agente causal de los daños. Las plantas presentaban afectaciones en las vainas, tallo y follaje iniciadas por insecto seguida por síntomas típicos de hongos fitopatógenos. Las muestras fueron procesadas y se determinó que el agente causal es el hongo fitopatógeno Fusarium semitectum. Este hongo se asocia a Fusicladium sp causando daños severos en la planta. En Cuba se ha aislado fundamentalmente en lotes de semillas de moringa.

It analyzed samples of Moringa oleifera from Habana province, Cuba with the objective of to determinate the causal agent of the damages. The plants shows affectations in pods, stems and foliage initiated for insect straight on typical symptoms of phytopathongens fungi. The samples were preprocessed and it determined that causal agent is phytopathongens fungus Fusarium semitectum. This fungus is associated to Fusicladium sp caused severe damages in the plant. In Cuba it has isolated principally from fractions of moringa seeds.

Arbuscular mycorrhizal fungi affect glucosinolate and mineral element composition in leaves of Moringa oleifera.

Moringa is a mycorrhizal crop cultivated in the tropics and subtropics and appreciated for its nutritive and health-promoting value. As well as improving plant mineral nutrition, arbuscular mycorrhizal fungi (AMF) can affect plant synthesis of compounds bioactive against chronic diseases in humans. Rhizophagus intraradices and Funneliformis mosseae were used in a full factorial experiment to investigate the impact of AMF on the accumulation of glucosinolates, flavonoids, phenolic acids, carotenoids, and mineral elements in moringa leaves. Levels of glucosinolates were enhanced, flavonoids and phenolic acids were not affected, levels of carotenoids (including provitamin A) were species-specifically reduced, and mineral elements were affected differently, with only Cu and Zn being increased by the AMF. This study presents novel results on AMF effects on glucosinolates in leaves and supports conclusions that the impacts of these fungi on microelement concentrations in edible plants are species dependent. The nonspecific positive effects on glucosinolates and the species-specific negative effects on carotenoids encourage research on other AMF species to achieve general benefits on bioactive compounds in moringa.

Bioactive secondary metabolites from Nigrospora sp. LLGLM003, an endophytic fungus of the medicinal plant Moringa oleifera Lam.

An endophytic fungus was isolated from the root of the medicinal plant Moringa oleifera Lam. Based on analyzing the rDNA sequence, the fungus was identified as Nigrospora sp. This is the first report of the isolation of endophytic Nigrospora from M. oleifera. By bioassay-guided fractionation, four antifungal secondary metabolites were isolated from liquid cultures of the fungus Nigrospora sp. LLGLM003, and their chemical structures were determined to be griseofulvin (1), dechlorogriseofulvin (2), 8-dihydroramulosin (3) and mellein (4) on the basis of spectroscopic analyses. Compound 2, 3 and 4 were isolated from Nigrospora sp. for the first time. In vitro antifungal assay showed that griseofulvin displayed clear inhibition of the growth of 8 plant pathogenic fungi. Dechlorogriseofulvin and mellein exhibited only weak antifungal activities, whereas 8-dihydroramulosin displayed no antifungal activities.

Development of a locally sustainable functional food for people living with HIV in Sub-Saharan Africa: laboratory testing and sensory evaluation.

The use of Lactobacillus rhamnosus GR-1 and micronutrients has been associated with a preserved immune function among people living with HIV. However, use of these products in the developing world remains limited due to the lack of facilities for production. We describe the development of a yogurt with L. rhamnosus GR-1 at >7×10(7) colony forming units fortified with locally grown Moringa oleifera leaves at 20% of the recommended daily allowance of vitamin A. The product was made by preparing a thin paste of Moringa which was then incubated with 4% probiotic and 2% yogurt mother culture in milk for 6 hours. The addition of M. oleifera enhanced the survival of probiotic bacteria in yogurt during the shelf life period at 5 °C (P=0.02), but had no effect on probiotic survival at 21 °C. While the sensory characteristics of probiotic and non-probiotic supplemented Moringa yogurts were indistinguishable, the addition of Moringa reduced consumer acceptance compared to regular yogurt.